Caulobacter FliQ and FliR membrane proteins, required for flagellar biogenesis and cell division, belong to a family of virulence factor export proteins

The Caulobacter crescentus fliQ and fliR genes encode membrane proteins that have a role in an early step of flagellar biogenesis and belong to a family of proteins implicated in the export of virulence factors. These include the MopD and MopE proteins from Erwinia carotovora, the Spa9 and Spa29 proteins from Shigella flexneri, and the YscS protein from Yersinia pestis. Inclusion in this family of proteins suggests that FliQ and FliR may participate in an export pathway required for flagellum assembly. In addition, mutations in either fliQ or fliR exhibit defects in cell division and thus may participate directly or indirectly in the division process. fliQ and fliR are class II flagellar genes residing near the top of the regulatory hierarchy that determines the order of flagellar gene transcription. The promoter sequence of the fliQR operon differs from most known bacterial promoter sequences but is similar to other Caulobacter class II flagellar gene promoter sequences. The conserved nucleotides in the promoter region are clustered in the -10, -20 to -30, and -35 regions. The importance of the conserved bases for promoter activity was demonstrated by mutational analysis. Transcription of the fliQR operon is initiated at a specific time in the cell cycle, and deletion analysis revealed that the minimal sequence required for transcriptional activation resides within 59 bp of the start site.

[1]  L. Shapiro,et al.  Genetic analysis of a temporally transcribed chemotaxis gene cluster in Caulobacter crescentus. , 1991, Genetics.

[2]  J. Wingrove,et al.  A sigma 54 transcriptional activator also functions as a pole-specific repressor in Caulobacter. , 1994, Genes & development.

[3]  T. Joys The covalent structure of the phase-1 flagellar filament protein of Salmonella typhimurium and its comparison with other flagellins. , 1985, The Journal of biological chemistry.

[4]  N. Ohta,et al.  Genetic switching in the flagellar gene hierarchy of Caulobacter requires negative as well as positive regulation of transcription. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[5]  P. Sansonetti,et al.  Localization of plasmid loci necessary for the entry of Shigella flexneri into HeLa cells, and characterization of one locus encoding four immunogenic polypeptides. , 1987, Journal of general microbiology.

[6]  R. Macnab,et al.  Genetic evidence for a switching and energy-transducing complex in the flagellar motor of Salmonella typhimurium , 1986, Journal of bacteriology.

[7]  J. Smit,et al.  Transformation of freshwater and marine caulobacters by electroporation , 1991, Journal of bacteriology.

[8]  G. Salmond,et al.  A pleiotropic reduced virulence (Rvi‐) mutant of Erwinia carotovora subspecies atroseptica is defective in flagella assembly proteins that are conserved in plant and animal bacterial pathogens , 1993, Molecular microbiology.

[9]  M. Venkatesan,et al.  Characterization of invasion plasmid antigen genes (ipaBCD) from Shigella flexneri. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[10]  F. Khambaty,et al.  Molecular genetics of the flgI region and its role in flagellum biosynthesis in Caulobacter crescentus , 1992, Journal of bacteriology.

[11]  L. Shapiro,et al.  A developmentally regulated Caulobacter flagellar promoter is activated by 3' enhancer and IHF binding elements. , 1992, Molecular biology of the cell.

[12]  R M Macnab,et al.  Genetics and biogenesis of bacterial flagella. , 1992, Annual review of genetics.

[13]  K. Namba,et al.  Morphological pathway of flagellar assembly in Salmonella typhimurium. , 1992, Journal of molecular biology.

[14]  C. Sasakawa,et al.  Eight genes in region 5 that form an operon are essential for invasion of epithelial cells by Shigella flexneri 2a , 1993, Journal of bacteriology.

[15]  J. Devereux,et al.  A comprehensive set of sequence analysis programs for the VAX , 1984, Nucleic Acids Res..

[16]  Y. Komeda,et al.  Incomplete flagellar structures in Escherichia coli mutants , 1981, Journal of bacteriology.

[17]  L. Shapiro,et al.  Identification of a Caulobacter basal body structural gene and a cis-acting site required for activation of transcription , 1990, Journal of bacteriology.

[18]  A. Newton,et al.  Regulation of the cell division cycle and differentiation in bacteria. , 1990, Annual review of microbiology.

[19]  L. Shapiro,et al.  Early Caulobacter crescentus genes fliL and fliM are required for flagellar gene expression and normal cell division , 1992, Journal of bacteriology.

[20]  M. Venkatesan,et al.  Surface presentation of Shigella flexneri invasion plasmid antigens requires the products of the spa locus , 1992, Journal of bacteriology.

[21]  J. Wingrove,et al.  Spatial and temporal phosphorylation of a transcriptional activator regulates pole-specific gene expression in Caulobacter. , 1993, Genes & development.

[22]  L. Shapiro,et al.  Caulobacter flagellar function, but not assembly, requires FliL, a non-polarly localized membrane protein present in all cell types. , 1994, Journal of molecular biology.

[23]  R. Macnab,et al.  Flagellar hook and hook-associated proteins of Salmonella typhimurium and their relationship to other axial components of the flagellum. , 1990, Journal of molecular biology.

[24]  D. DeRosier,et al.  Size of the export channel in the flagellar filament of Salmonella typhimurium. , 1993, Ultramicroscopy.

[25]  R. Brasseur,et al.  Secretion of Yop proteins by Yersiniae , 1990, Infection and immunity.

[26]  L. Shapiro,et al.  Genetic regulatory hierarchy in Caulobacter development. , 1990, Advances in genetics.

[27]  P. Matsumura,et al.  DNA sequence analysis, gene product identification, and localization of flagellar motor components of Escherichia coli , 1989, Journal of bacteriology.

[28]  J. Poindexter BIOLOGICAL PROPERTIES AND CLASSIFICATION OF THE CAULOBACTER GROUP , 1964, Bacteriological reviews.

[29]  L. Shapiro,et al.  Cascade regulation of Caulobacter flagellar and chemotaxis genes. , 1987, Journal of molecular biology.

[30]  R. Macnab,et al.  Flagellar assembly in Salmonella typhimurium: analysis with temperature-sensitive mutants , 1990, Journal of bacteriology.

[31]  Jeffrey H. Miller Experiments in molecular genetics , 1972 .

[32]  L. Shapiro,et al.  Expression of an early gene in the flagellar regulatory hierarchy is sensitive to an interruption in DNA replication , 1992, Journal of bacteriology.

[33]  L. Shapiro,et al.  Identification of a gene cluster involved in flagellar basal body biogenesis in Caulobacter crescentus. , 1987, Journal of molecular biology.

[34]  N. Agabian,et al.  Envelope-associated nucleoid from Caulobacter crescentus stalked and swarmer cells , 1977, Journal of bacteriology.

[35]  L. Shapiro,et al.  A developmentally regulated chromosomal origin of replication uses essential transcription elements. , 1995, Genes & development.

[36]  L. Shapiro,et al.  An unusual promoter controls cell‐cycle regulation and dependence on DNA replication of the Caulobacter fliLM early flagellar operon , 1993, Molecular microbiology.

[37]  R. Macnab,et al.  Export of an N-terminal fragment of Escherichia coli flagellin by a flagellum-specific pathway. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[38]  R. Macnab,et al.  L-, P-, and M-ring proteins of the flagellar basal body of Salmonella typhimurium: gene sequences and deduced protein sequences , 1989, Journal of bacteriology.

[39]  R M Macnab,et al.  Salmonella typhimurium mutants defective in flagellar filament regrowth and sequence similarity of FliI to F0F1, vacuolar, and archaebacterial ATPase subunits , 1991, Journal of bacteriology.

[40]  T. Suzuki,et al.  Incomplete flagellar structures in nonflagellate mutants of Salmonella typhimurium , 1978, Journal of bacteriology.

[41]  K. Namba,et al.  Structure of the core and central channel of bacterial flagella , 1989, Nature.

[42]  L. Shapiro,et al.  A temporally controlled sigma-factor is required for polar morphogenesis and normal cell division in Caulobacter. , 1992, Genes & development.

[43]  L. Shapiro,et al.  Organization and temporal expression of a flagellar basal body gene in Caulobacter crescentus , 1988, Journal of bacteriology.

[44]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[45]  G. Ordal,et al.  Nucleotide sequences of Bacillus subtilis flagellar biosynthetic genes fliP and fliQ and identification of a novel flagellar gene, fliZ , 1992, Journal of bacteriology.

[46]  L. Shapiro,et al.  Coordinate cell cycle control of a Caulobacter DNA methyltransferase and the flagellar genetic hierarchy , 1995, Journal of bacteriology.

[47]  G. Ordal,et al.  Bacillus subtilis flagellar proteins FliP, FliQ, FliR and FlhB are related to Shigella flexneri virulence factors. , 1993, Gene.

[48]  A. Ninfa,et al.  The Caulobacter crescentus FlbD protein acts at ftr sequence elements both to activate and to repress transcription of cell cycle-regulated flagellar genes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[49]  L. Shapiro,et al.  Organization and ordered expression of Caulobacter genes encoding flagellar basal body rod and ring proteins. , 1992, Journal of molecular biology.

[50]  A. Newton,et al.  Identification of the promoter and a negative regulatory element, ftr4, that is needed for cell cycle timing of fliF operon expression in Caulobacter crescentus , 1993, Journal of bacteriology.

[51]  L. Shapiro,et al.  Methylation involved in chemotaxis is regulated during Caulobacter differentiation. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[52]  L. Shapiro,et al.  Negative transcriptional regulation in the Caulobacter flagellar hierarchy. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[53]  G. Salmond,et al.  A pleiotropic reduced virulence (Rvi−) mutant of Erwinia carotovora subspecies atroseptica is defective in flagella assembly proteins that are conserved in plant and animal bacterial pathogens , 1993, Molecular microbiology.

[54]  A. Newton,et al.  FlbD of Caulobacter crescentus is a homologue of the NtrC (NRI) protein and activates sigma 54-dependent flagellar gene promoters. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[55]  P. Matsumura,et al.  Molecular characterization, nucleotide sequence, and expression of the fliO, fliP, fliQ, and fliR genes of Escherichia coli , 1994, Journal of bacteriology.

[56]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[57]  T. Bergman,et al.  The lcrB (yscN/U) gene cluster of Yersinia pseudotuberculosis is involved in Yop secretion and shows high homology to the spa gene clusters of Shigella flexneri and Salmonella typhimurium , 1994, Journal of bacteriology.

[58]  R. Macnab,et al.  The flaFIX gene product of Salmonella typhimurium is a flagellar basal body component with a signal peptide for export , 1987, Journal of bacteriology.

[59]  K. Hughes,et al.  Sensing structural intermediates in bacterial flagellar assembly by export of a negative regulator. , 1993, Science.

[60]  R. Macnab,et al.  Identification of flagellar hook and basal body gene products (FlaFV, FlaFVI, FlaFVII and FlaFVIII) in Salmonella typhimurium , 1987, Journal of bacteriology.

[61]  R. Macnab,et al.  Subdivision of flagellar genes of Salmonella typhimurium into regions responsible for assembly, rotation, and switching , 1986, Journal of bacteriology.

[62]  H. Wolf‐Watz,et al.  The virulence protein Yop5 of Yersinia pseudotuberculosis is regulated at transcriptional level by plasmid‐plB1 ‐encoded trans‐acting elements controlled by temperature and calcium , 1988, Molecular microbiology.

[63]  J. Rine,et al.  Transmission electron microscopy and immunocytochemical studies of yeast: analysis of HMG-CoA reductase overproduction by electron microscopy. , 1989, Methods in cell biology.

[64]  R. C. Johnson,et al.  Analysis of nonmotile mutants of the dimorphic bacterium Caulobacter crescentus , 1979, Journal of bacteriology.